Robotic Production of Cancer Cell Spheroids with an Aqueous Two-phase System for Drug Testing
Robotic Production of Cancer Cell Spheroids with an Aqueous Two-phase System for Drug Testing
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DOI：
10.3791/52754
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发表时间：
2015-04-01
影响因子：
1.2
通讯作者：
Tavana, Hossein
中科院分区：
文献类型：
--
作者：
Ham, Stephanie Lemmo;Atefi, Ehsan;Tavana, Hossein
癌细胞球体为抗癌药物测试应用提供了相关的无血管肿瘤体外模型。这项工作描述了以高通量 96 孔板形式生产单一培养和共培养球体的详细协议。该方法利用水性两相系统将细胞限制在浸没在第二水相中的较致密水相的液滴中。液滴停留在孔表面上，使细胞紧密靠近，形成单个球体。该技术已适用于机器人液体处理机，以生产尺寸受控的球体，并加快用于化合物筛选应用的球体生产过程。用临床使用的药物处理的球体显示，随着药物剂量的增加，细胞活力降低。使用标准微孔板生成球体，可以使用酶标仪轻松分析经过药物处理的球体的癌细胞的活力。该技术可以直接通过机器人和其他液体处理工具（例如手动移液器）实施。
Cancer cell spheroids present a relevant in vitro model of avascular tumors for anti-cancer drug testing applications. A detailed protocol for producing both mono-culture and co-culture spheroids in a high throughput 96-well plate format is described in this work. This approach utilizes an aqueous two-phase system to confine cells into a drop of the denser aqueous phase immersed within the second aqueous phase. The drop rests on the well surface and keeps cells in close proximity to form a single spheroid. This technology has been adapted to a robotic liquid handler to produce size-controlled spheroids and expedite the process of spheroid production for compound screening applications. Spheroids treated with a clinically-used drug show reduced cell viability with increase in the drug dose. The use of a standard micro-well plate for spheroid generation makes it straightforward to analyze viability of cancer cells of drug-treated spheroids with a micro-plate reader. This technology is straightforward to implement both robotically and with other liquid handling tools such as manual pipettes.